JP2005162685A - Antithrombotic composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain an antithrombotic composition widely usable for food and drink; and to provide the food and drink containing the antithrombotic composition. <P>SOLUTION: This antithrombotic composition contains an extract of tea. Preferably, the extract of the tea is the tea extract containing a reduced amount of polyphenols. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to an antithrombotic composition containing a tea extract and a food or drink containing the composition.

A thrombus can be formed into an insoluble polymer together with platelets, leukocytes and the like and solidified on the inner wall of a blood vessel while a protein called fibrinogen is activated and converted into fibrin. When the body is normal, the fibrinolytic enzyme that works to dissolve fibrin, which is the source of the thrombus, prevents thrombus. If the fibrinolytic enzyme is insufficient, fibrin cannot be dissolved and a thrombus can be formed.
The formed thrombus is deposited in the blood vessels, reducing the cross-sectional area of the blood vessels and impeding blood circulation, so that the blood cannot normally supply nutrients and oxygen in cells and tissues, Problems such as the inability to discharge cell and tissue waste and accumulation of toxicity will occur.
In blood vessels, the symptoms caused by blood clots called blood clots are called thrombosis in a broad sense (hereinafter simply referred to as “thrombosis” in the broad sense) and are caused by blood clots. The pathological condition is divided into thrombosis and embolism in a narrow sense. In the narrow sense, thrombosis is a symptom caused by partial or complete blockage of blood flow at the site where the thrombus is formed, and embolism is caused by removal of the thrombus from the formation site and movement by the blood flow. It refers to a pathological condition caused by physical or complete occlusion.
Such thrombosis induces various diseases depending on the site of the blood vessel where the thrombus has occurred. In particular, serious symptoms such as stroke, cerebral hemorrhage, cerebral infarction, heart failure, myocardial infarction, heart palsy occur when it occurs in the cerebral blood vessel or cardiovascular, causing half-body involuntary and death in severe cases is there.

Currently, in order to solve thrombosis, research and development of antithrombotic agents and thrombus formation preventive agents that suppress the formation of thrombus and thrombolytic agents that dissolve the generated thrombus are mainly conducted.
As an antithrombotic agent or an agent for preventing thrombus formation, aspirin that inhibits blood coagulation by inhibiting the adhesion of platelets to the blood vessel wall, heparin (Heparin), coumarin (Coumarin) that blocks the body's intrinsic blood coagulation pathway ) Etc. are currently used in clinical practice. Recently, eicosapentaenoic acid (EPA), prostacycline (PG12) derivatives and the like have been commercialized. However, since these drugs have no specificity, they may affect parts other than the thrombus in the living body, and may cause bleeding or the like when remaining in the living body. In addition, antithrombotic activities such as hirudin, synthetic antithrombin, and ticlopidin have been reported, but have not yet been put into practical use.
As a thrombolytic agent, a plasminogen activator (plasminogen activator) such as streptokinase or urokinase is intravenously injected into a patient in which a thrombus has been generated to activate the thrombolytic system in the body. Is commonly used. The effect of dissolving thrombus has been proved in many clinical experiments, but, like antithrombotic agents or antithrombotic agents, it has no specificity for thrombus and has side effects such as general bleeding during the treatment of thrombus. is there. In addition, tissue-type plasminogen activator (tPA) was considered to be an ideal thrombolytic agent with high selectivity for thrombus, but as a result of actual application to clinical treatment, Although there were differences, there were still side effects such as systemic bleeding. In addition, since the half-life in the blood is very short and the duration of the drug is short, the dose must be large in order to maintain the drug in the body, so the treatment cost is much higher than that of conventional thrombolytic agents. There is a problem that it is expensive.

Although these medicines are used to prevent the formation of blood clots, they do not show a significant effect on thrombus removal and induce serious side effects. Research on ingredients or food ingredients having a function of preventing or regulating or activating the constitution has also attracted attention.
As food ingredients, materials such as nattokinase, polyunsaturated fatty acids, glucosamine, and onion skin (for example, see Patent Document 1) are known, but there are problems with flavor and properties, and they are widely applied to foods. could not.
In addition, recently, a patent for a kiwifruit extract (for example, see Patent Document 2) has been published, but there is a drawback that the activity in the neutral range is weak.

JP 2002-171934 A (2nd page) JP 2003-171294 A (page 2-5)

  An object of the present invention is to provide an antithrombotic composition that can be used for a wide range of foods and drinks and foods and drinks containing the same.

  As a result of diversified research and studies for the purpose of searching for antithrombotic components using various natural plants, the present inventors have found that tea extract has an excellent antithrombotic effect, and completed the present invention. It was.

The antithrombotic composition containing the tea extract obtained in the present invention inactivates factors involved in the intrinsic pathway from the result of measuring the activated partial thromboplastin time (APTT). It was found that the effect of inhibiting fibrin formation and suppressing thrombus formation in blood vessels is high.
In particular, tea is safe because it has no side effects that cause bleeding in the body, unlike conventional drugs because it is derived from a natural plant that humans have been using for a long time.
The present invention uses an antithrombotic composition containing a tea extract for various foods and beverages, pharmaceuticals, and the like, and suppresses the formation of thrombus, thereby causing cerebral hemorrhage, cerebral infarction, myocardial infarction, arteriosclerosis and coronary artery disease. Can prevent various cardiovascular diseases.

  The tea used in the present invention is not particularly limited, and botanically includes green tea that is non-fermented tea produced from leaves of the camellia plant, oolong tea that is semi-fermented tea, and black tea that is fermented tea. It is done. Among them, it is preferable to use green tea which is non-fermented tea from the viewpoint of effect.

From the viewpoint of flavor, the tea extract of the present invention is preferably reduced in polyphenols which are astringent ingredients.
As a method for removing polyphenol which is an astringent component, a method of adding polyvinyl polypyrrolidone to an extract (Japanese Patent Laid-Open No. 1-218550) or a method of removing polyphenol together with water dehydrated by freeze concentration (Japanese Patent Laid-Open No. 2002-2002). 325539) etc. are generally known and available.
The method for measuring the polyphenol content in the present invention is not particularly limited, and examples thereof include an iron tartrate colorimetric method and a foreign thiocult method, and an iron tartrate colorimetric method is preferable. The polyphenol content is preferably 15% or less, more preferably 10% or less in the solid content of the tea extract from the aspect of taste.

  Here, for example, in the case of collecting polyphenols, the residue after extracting the polyphenols from the tea water or the hot water extract is exactly the “tea with reduced polyphenols” of the present invention. Corresponds to “extract”. In addition, since polyphenols have the property of being easily dissolved in a specific organic solvent, for example, when fractionation is performed using a specific organic solvent, most of them are dissolved on the organic solvent side (water-insoluble fraction). However, it is hardly contained in the water fraction. That is, it is obtained from a water transfer fraction obtained by extracting tea leaves or crushed tea leaves with water or hot water and distributing them to a solvent such as ethyl acetate or acetone. In recent years, in view of various functionalities possessed by polyphenols, the utilization of polyphenols has increased, and the components after extraction of polyphenols from hot water extract of tea are so-called by-products (or residues). And was discarded without being effectively used. In that respect, according to the present invention, the components that have been discarded without being effectively used so far can be used effectively, so that the hot water extract of tea can be used without any waste, improving economic efficiency and disposal. There is also an advantage that it leads to a reduction in the amount of waste discharged.

In addition, the tea extract of the present invention preferably has reduced caffeine, which has a function of increasing the blood concentration of prothrombin and fibrinogen that cause clotting.
As a method for removing caffeine, a method using acidic clay (Japanese Patent Laid-Open No. 06-142405), a method using activated carbon (Japanese Patent Laid-Open No. 08-077072), and a method using a synthetic adsorbent (Japanese Patent Laid-Open No. 08-109178). Etc.) are generally known and available.
The method for measuring caffeine is not particularly limited, and examples thereof include high performance liquid chromatography. The caffeine content is preferably 2% or less, more preferably 1% or less in the solid content of the tea extract.

  The method of separating the hot water extract of tea from the water fraction of the distribution product with ethyl acetate or acetone is preferable because caffeine can be reduced simultaneously with polyphenol.

  Further, in the above extraction, it is preferable to repeat the extraction step once or more for the extraction residue, since the extraction rate is improved. The solvent used for extraction in this case may be the same, or another solvent may be used.

Although the above extract can be used as it is, it is preferable because the antithrombotic effect is enhanced and the application range is widened by removing insoluble substances by filtration or centrifugation.
What removed the insoluble substance and then recovered the precipitate obtained by adding ethanol to the extract is preferable because the antithrombotic effect is further enhanced. Although it does not specifically limit as a density | concentration of ethanol, 10-50% is preferable as a final concentration from the point of a yield and an effect, and 15-45% is still more preferable.
The extract can be used as it is, but it can also be dried if necessary.

The antithrombosis in the present invention is not particularly limited, but is preferably an action (anticoagulation action) that suppresses the formation of a thrombus.
Although the measurement of the antithrombotic effect in the present invention is not particularly limited, for example, there is a method for measuring the anticoagulant activity against the endogenous blood coagulation system, and the activated partial thromboplastin time (APTT) is measured. This can be confirmed.

The antithrombotic composition of the present invention can be applied to foods and drinks, pharmaceuticals, feeds, and the like, and preferably foods and drinks that can be easily consumed by humans.
The food and drink in the present invention is not particularly limited as long as it is a form that can be taken orally, such as a solution, suspension, powder, or solid molded product. More specifically, instant noodles, retort foods, canned foods, microwave foods, instant soups and miso soups, freeze-dried foods, soft drinks, fruit juice drinks, vegetable drinks, soy milk drinks, coffee drinks, tea drinks Beverages such as powdered beverages, concentrated beverages, nutritional beverages, alcoholic beverages, bread, pasta, noodles, cake mixes, flour products such as fried flour, bread crumbs, rice cakes, caramel, chewing gum, chocolate, cookies, biscuits, cakes, Sweets such as pies, snacks, crackers, Japanese sweets, desserts, sauces, processed tomato seasonings, flavor seasonings, cooking mixes, sauces, dressings, soups, curry and stew seasonings, processing Fats and oils such as butter, margarine and mayonnaise, milk beverages, yogurts, lactic acid bacteria beverages, ice creams, chestnuts Agricultural processing such as dairy products such as mussels, frozen foods, processed fishery products such as fish meat ham and sausage, fish paste products, livestock processed products such as livestock ham and sausage, canned agricultural products, jams and marmalades, pickles, boiled beans and cereals Products, nutritional foods, tablets, capsules and the like.

In the present invention, various nutritional components can be strengthened when processed into an antithrombotic composition or a food or drink.
Nutritional ingredients that can be enhanced include vitamins such as vitamin A, vitamin B ₁ , vitamin B ₂ , vitamin B ₆ , vitamin B ₁₂ , vitamin C, vitamin D, vitamin E, niacin (nicotinic acid), pantothenic acid, folic acid, Essential amino acids such as lysine, threonine and tryptophan, minerals such as calcium, magnesium, iron, zinc and copper, and for example, α-linolenic acid, EPA, DHA, evening primrose oil, octacosanol, casein phosphopeptide (CPP), Casein calcium peptide (CCP), water-soluble dietary fiber, insoluble dietary fiber, substances that contribute to human health, such as oligosaccharides, and one or more useful substances approved as other foods and food additives Can be used.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, the following Example does not limit the scope of the present invention.

(Example 1) Preparation 1 of antithrombotic composition
Add 15 kg of hot water to 1 kg of green tea leaf, extract at 90 ° C. for 30 minutes, filter to remove the tea husk, add 12 kg of ethyl acetate to the filtrate, shake, stand, and distribute. . The water fraction was collected, desolvated under reduced pressure (0.067 mpa), and spray-dried to obtain 110 g of anti-thrombotic composition A of the present invention.
This had a polyphenol content of 9.2% and a caffeine content of 0.7%.

(Test Example 1) Confirmation of antithrombotic effect The anticoagulant activity of the antithrombotic composition of the present invention was measured with a coagulometer using platelet poor plasma (Platelet Poor Plasma, PPP) separated from human blood. It was measured.
In a reaction cuvette, 10 microliters of sample, 25 microliters of APTT reagent, and 25 microliters of 10% Sephaline were added and reacted at 37 ° C. for 3 minutes, and then 50 microliters of PPP was added and reacted for 2 minutes. Finally, 50 microliters of 25 mM calcium chloride was added and allowed to clot, and the time until the plasma was clotted was measured and designated APTT.
At this time, water was added to the control and APTT was measured by the same method. The anticoagulant activity (%) relative to the control was calculated from the measured APTT of the sample by the following formula.
Anticoagulant activity (%) = ((sample APTT−control APTT) / control APTT) × 100
In order to confirm the relationship between the concentration of the antithrombotic composition of the present invention and the anticoagulant activity, the antithrombotic composition of the present invention of 0 (control), 1, 3, 5 mg / ml as the solid content concentration Was used to measure its activity. The results are shown in Table 1 below.

Table 1

  From the results of Table 1 above, it was confirmed that the antithrombotic composition of the present invention showed high anticoagulant activity. It was also confirmed that the anticoagulant activity increased proportionally by increasing the extract concentration.

(Example 2) Preparation 2 of antithrombotic composition
To 100 grams of the antithrombotic composition A obtained in Example 1, 2 liters of water was added to dissolve, ethanol was added to prepare 2.22 liters (final ethanol concentration 20%), and then at room temperature. It was allowed to stand for 24 hours to precipitate insoluble components. The precipitate was separated by centrifugation, dried under reduced pressure, redissolved in 2 liters of water, filtered to remove insoluble components, the filtrate was freeze-dried, and 25 g (yield) of anti-thrombotic composition B of the present invention was obtained. 25%).
Similarly, the final concentrations of ethanol were 40%, 60%, and 80% to obtain antithrombotic compositions C, D, and E of the present invention.
The polyphenol contents of the antithrombotic compositions B to E obtained here are B: 1.1%, C: 1.5%, D: 1.8%, E: 2.3%, respectively. The in contents were B: 0.1%, C: 0.3%, D: 0.4%, and E: 0.5%, respectively.

(Test Example 2) Confirmation of antithrombotic effect The antithrombotic compositions BE obtained in Example 2 were measured for APTT at a concentration of 5 mg / ml. The results are shown in Table 2.

Table 2

  From the results in Table 2 above, it was found that the antithrombotic effect was highest when the ethanol concentration was 40%.

(Example 3) Preparation of antithrombotic composition-containing food (tablet confectionery) 50 g of antithrombotic composition B obtained in Example 2, 30 g of lactose, DHA-containing powdered oil (Suncoat DY-5; Taiyo Chemical Co., Ltd.) 12 g, 4 g of sucrose fatty acid ester and 4 g of yoghurt flavor were mixed and tableted so that 1 tablet would be 300 mg to obtain a food / beverage product (tablet cake) containing the antithrombotic composition of the present invention.

Example 4 Preparation of Antithrombotic Composition-Containing Beverage (Vegetable Juice Mixed Beverage) 1 g of antithrombotic composition B obtained in Example 2 and guar gum degradation product (Sunfiber R; manufactured by Taiyo Chemical Co., Ltd.) 3 g was added to, mixed and dissolved in 100 ml of a commercially available vegetable juice mixed beverage to obtain a food / beverage product containing the composition for antithrombosis of the present invention (vegetable juice mixed beverage).

The embodiment of the present invention and the target product are as follows.
(1) An antithrombotic composition comprising a tea extract.
(2) The antithrombotic composition as described in (1) above, wherein the tea extract is a tea extract with reduced polyphenols.
(3) The antithrombotic composition as described in (1) or (2) above, wherein the content of polyphenols in the tea extract is 15% or less in the solid content of the tea extract.
(4) The antithrombotic composition according to any one of (1) to (3) above, wherein the content of polyphenols in the tea extract is 10% or less in the solid content of the tea extract.
(5) The antithrombotic composition according to (1) to (4) above, wherein the tea extract is a tea extract with reduced caffeine.
(6) The antithrombotic composition as described in any of (1) to (5) above, wherein the content of caffeine in the tea extract is 2% or less in the solid content of the tea extract.
(7) The antithrombotic composition according to any one of (1) to (6) above, wherein the content of caffeine in the tea extract is 1% or less in the solid content of the tea extract.
(8) The above-mentioned (2) to (7), wherein the tea extract with reduced polyphenols and / or caffeine is a water fraction of a distribution product of hot water extract of tea with ethyl acetate. Any antithrombotic composition.

(9) The antithrombosis according to any one of (1) to (8) above, wherein the tea extract is further fractionated with ethanol from a tea extract with reduced polyphenols and / or caffeine Composition.
(10) The ethanol concentration according to any one of (1) to (9) above, wherein the ethanol concentration at the time of fractionation with ethanol is 10 to 50% as a final concentration and is a precipitate fraction thereof Composition.
(11) Antithrombotic use according to any one of (1) to (10) above, wherein the ethanol concentration at the time of fractionation with ethanol is 20 to 40% as a final concentration and is a precipitate fraction thereof Composition.
(12) A food or drink comprising the antithrombotic composition according to any one of (1) to (11).
(13) A pharmaceutical comprising the antithrombotic composition according to any one of (1) to (11).
(14) A feed comprising the antithrombotic composition according to any one of (1) to (11).

  The antithrombotic composition containing tea extract obtained in the present invention inhibits the activity of thrombin which forms fibrin which is the final stage with many enzymes involved in the intrinsic pathway of blood coagulation system. It has a high antithrombotic effect that suppresses thrombus formation, and is used in various foods and medicines and medicines to suppress the formation of thrombus, thereby preventing heart blood such as cerebral hemorrhage, cerebral infarction, myocardial infarction, arteriosclerosis and coronary artery disease. Related diseases can be prevented.

Claims (5)

An antithrombotic composition comprising a tea extract. The antithrombotic composition according to claim 1, wherein the tea extract is a tea extract with reduced polyphenols. 3. The antithrombotic composition according to claim 1 or 2, wherein the tea extract is a tea extract with reduced caffeine. The antithrombotic agent according to any one of claims 1 to 3, wherein the tea extract is a precipitate obtained by further fractionating the tea extract with reduced polyphenols and / or caffeine with ethanol. Composition. A food or drink comprising the antithrombotic composition according to any one of claims 1 to 4.